Figure 8.

mRNA and protein expressions of factors involved in regulation of metabolism in myotubes cultured in a lactic acid-containing medium. Myotubes were cultured in standard cell media (basal, 5.5 mM glucose) or cell media where glucose had been replaced with lactic acid (5 mM) for the whole proliferation and differentiation period. After ended differentiation, total cell RNA and protein contents were isolated. (a) mRNA expressions of CD36 molecule (CD36), solute carrier family 2 member 4 (GLUT4), angiopoietin like 4 (ANGPTL4), pyruvate dehydrogenase kinase 4 (PDK4), carnitine palmitoyl transferase 1 (CPT1A and B) and cytochrome c1 (CYC1). All values were corrected for the housekeeping control RPLP0, and presented as means ± SEM relative to basal (n = 5). (b,c) Expressions of proteins involved in mitochondrial phosphorylation, OXPHOS (complex I subunit NDUFB8, complex II subunit, complex III subunit core 2, complex IV subunit II and complex V (ATP synthase subunit α)). (b) One representative immunoblot. (c) Quantified immunoblots normalized to glucose. All values were corrected for the housekeeping control α-tubulin, normalized to glucose, and data are presented as means ± SEM relative to basal (n = 5). Dividing lines delineate blots from different gels. The samples derive from the same experiment, and all blots were processed in parallell. Full-length blots are presented in Supplementary Fig. 8. *Statistically significant vs. basal (p < 0.05, paired Student’s t-test).